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CHAPTER

26
Diagnostic Value of

Necropsy
MADELINE A. RAE, BS, DVM, MS, D ipl ABVP-A vian

Gross necropsy and postmortem diagnostic testing are

important parts of avian medicine, requiring a systematic
approach to the examination of organs and the collec-
tion of samples.

Necropsy examination functions as more than a way to

satisfy the curiosity of the client, breeder or attending
veterinarian — it provides important information that
can be used in the diagnosis and treatment of future
cases. Clinical signs and clinical pathologic findings are
often not definitively explained until necropsy.

Postmortem information can be invaluable in educating

clients regarding the seriousness of husbandry, nutri-
tional and infectious disease conditions, thereby pre-
venting them from making the same mistakes with sub-
sequent birds. For grieving owners, necropsy findings
can relieve them of some or all of the guilt associated
with the death of a beloved pet.

Necropsy findings are an integral part of the flock data-

base from which husbandry, management, treatment,
vaccination and quarantine recommendations can be
made.

In situations where the client may be dissatisfied with

treatment or outcomes, it is wise to have a veterinary
pathologist perform the necropsy. Developing a relation-
ship with a veterinary pathologist is also highly recom-
mended so that appropriate samples are submitted, opti-
mizing the chances of arriving at a diagnosis. The veteri-
nary pathologist should have training, experience and
interest in companion and free-ranging birds.
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Preparing for the Necropsy
The supplies needed for an avian necropsy are listed in
Table 26.1. Although bottles or jars of neutral buffered
10% formalin should be available for the collection of
specimens for histopathology, some cautions are in
order. Ensure that formalin fumes do not contact tissues
that are to be cultured for bacteria or viruses, as this can
compromise the culture accuracy. Make certain that for-
malin fumes do not come in contact with blood or tissue
cytological smears, as this can severely distort staining
and interpretation.
A standardized necropsy form should be used and diag-
nostic specimen accession forms and instructions should
be readily located. All specimen containers should be
clearly labeled with the appropriate information. A
refrigerator, a freezer, insulated shipping containers and
coolant packs should be available for appropriate han-
dling and shipping of specimens. After the necropsy has
been performed, the carcass can be frozen and saved for
a period of time, as frozen tissues may be useful if initial
testing is inconclusive. Have arrangements in place for
appropriate disposal of carcasses and infectious wastes.
Table 26.1 | Supplies Needed for Avian Necropsy
• Gram scale
• Apron
• Mask
• Gloves
• Disinfectant
• Necropsy checklist
• Scalpel handle and blades
• Scissors*
• Thumb forceps*
• Small rongeurs or toenail nippers for brain removal and
bone cutting
• Ophthalmic scissors and forceps for small passerines,
neonate and dead-in-shell
• Sturdy paper plates and/or plastic cutting board
• Microscope slides and coverslips
• Blood and fluid collection tubes
• Sterile saline
• Sterile culturettes (aerobic and anaerobic)
• Sterile cotton-tipped applicators
• Magnifying head loupe
• Good light source
• Sterile sealable plastic bags or sterile plastic vials for
sample collection
• Plastic screw-top jars containing 10% neutral buffered
formalin
• Sterile syringes and needles
• Stains (Gram’s, Wright’s, Giemsa or Diff-Quik, acid-fast)
• Butane lighter or other heat source to heat-fix smears
for acid-fast staining
*Autoclave one set of scissors and forceps for collecting samples
aseptically.
Page 662
METHOD OF EUTHANASIA
In some instances, it may be appropriate to euthanize a
sick bird in order to diagnose a flock problem. The
method of euthanasia is important to consider because
many injectable agents can cause severe artifacts in tis-
sues. If at all possible, barbiturates should not be
injected into the coelomic cavity, thoracic cavity or heart.
The barbiturates are very acidic and crystals readily pre-
cipitate, causing severe tissue destruction that may
obscure gross and histologic lesions. Even intravenous
barbiturate solutions can cause intravascular erythrocyte
lysis and some tissue damage as the solution pools
within blood-filled organs.
Gas anesthetic agents seem to provide the least amount
of tissue artifact (less muscle contraction artifact, no cell
lysis). Once the gas agents anesthetize the bird, there is
the opportunity to collect antemortem blood samples
for hematology, serum chemistries, hemoparasite detec-
tion, serology and toxicology. The gas anesthetic agents
can then be followed with intravenous euthanasia
agents, if needed, and the amount of injectable agent
necessary is usually quite reduced.
PREPARING THE BODY
The necropsy should be performed as soon after death
as is possible. In order to prevent dry feathers from insu-
lating the body and delaying cooling, wet the feathers
with a detergent and water solution. The detergent and
water also decreases the dispersal of feather and fecal
dust into the local environment, thereby decreasing the
transmission of infectious agents.
The body should be refrigerated, not frozen. Freezing
can create artifacts in the tissues that may seriously
obscure histologic lesions. Postmortem autolysis also can
obscure histologic lesions, so if necropsy cannot be per-
formed within 3 days, the body should be frozen, realiz-
ing that histopathology is likely to be compromised.
When shipping a cooled body, be sure the ice packs (or
other frozen coolants) do not directly touch the body as
this may freeze the body tissues, especially in very small
birds. Wrap the ice packs in bubble wrap or newspaper
to prevent freeze damage.
GUIDELINES FOR OBTAINING
THE HISTORY
A detailed history should be obtained, just as the clini-
cian would do upon seeing a live bird for the first time in
the examination room (Table 26.2) (see also Chapter 6,
Maximizing Information from the Physical Examination).
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Table 26.2 | Considerations for Obtaining History Information

• Start with the age of the bird and sex,
if known. Breeders are often very inter-
ested in having the sex of the dead bird
confirmed at necropsy. If the dead bird
is from a pair that is known to have
produced fertile eggs, the knowledge
of the mate can be deduced from the
confirmation of the dead bird’s sex. The
breeder then knows which sex of bird
to replace in the pair.
• Determine what the bird’s purpose is,
whether it is a dear pet, a display bird
or a breeder bird.
• It is important to obtain a detailed
description of the diet fed, and for how
long, including any supplements and
grit, brands used and their storage.
Keeping a stored sample frozen in an
airtight container may be useful.
Quality control by the manufacturer
and storage of formulated diets can be
a problem. The resulting products may
be over-formulated or rancid, resulting
in toxic levels over time.
• Determine if there have been changes
in appetite and/or droppings.
• How is water provided and how often
is it changed?
• Have feeding practices or the diet
changed recently?
• If this is a chick that is being hand-fed,
obtain a detailed description of the
feeding practices, brand of food with
amounts and times per day, as well as
any charts documenting weight loss
or gain.
• Is the bird allowed to fly freely within
the house?
• Ask for a description of the cage and
its placement within the house (eg,
near the kitchen where exposure to
toxic fumes may occur).
• Is bedding material used and how
often is the cage cleaned? What
products are used for cleaning?
• Are there toys in the cage and does or
can the bird chew on them?
• If this bird is from an aviary, a descrip-
tion of the aviary and floor plan can be
very helpful.
• How many other birds are in the home
or aviary and how many have died in
the last year? Are any other birds in
the household or aviary ill? Was the
deceased bird in quarantine? Are any
humans in contact with the ill birds?
• Have there been any recent changes in
environmental conditions such as tem-
perature changes?
• Have there been recent additions to
the aviary or household?
• Ask about the reproductive history of
the bird, such as a recent history of
egg laying, dystocia or feeding of
chicks.
• Obtain a description of recent clinical
signs noticed prior to death and their
duration.
• Ask about any other previous illnesses
or conditions. Were any medications or
treatments given to the bird? Be sure to
ask about prescription medications as
well as over-the-counter preparations.

PREVENTING CONTAMINATION It is important to collect samples of everything (all

Perform the necropsy in a well-lighted, well-ventilated
area (preferably under a fume hood), and wear gloves, a
mask and, if possible, a disposable apron. Aerosols from
feathers, feces and exudates can be infectious. This is
particularly important with cases of chlamydophilosis
and mycobacteriosis, which can be zoonotic. However, it
also is important to contain the feather dander and feces
in cases of avian polyomavirus and psittacine circovirus
infections, so as not to contaminate the premises, your
clothing or other adjacent birds.
organs, the grossly normal and abnormal). Labeling
sealable bags and formalin jars prior to the necropsy
with the owner’s name and the tissues enclosed can
save time and prevent interruptions in the flow of the
necropsy.
After the necropsy is completed, the decision of which
samples to send and what tests to request can be made,
and at the very least, the diagnosis will not be cremated
with the carcass.

Disinfectant solutions should be readily available for

clean-up after the necropsy, but neither these solutions
nor their fumes should come in contact with tissues
being collected, as they may lyse cells and destroy
microorganisms needed for culture.
Step-by-Step
Necropsy Procedure
The particular routine used for gross necropsy of birds
can vary, but what remains the same is that all organs
and systems are examined, and the use of a checklist will
ensure this. Use the Necropsy Checklist (Table 26.3) to
document all findings, both normal and abnormal. Make
this checklist a part of the medical record and send a
copy to the veterinary pathologist with any fixed tissues.
EXTERNAL EXAMINATION
The necropsy begins with an external examination.
Record the band number and scan for microchips; these
can be removed, labeled and saved as proof of identifica-
tion. Weigh the bird using a gram scale and record the
weight on the checklist. Palpate for obvious fractures;
radiographs may be warranted in some instances.
Examine the skin and feathers: often, feather abnormali-
ties may not be visible while the feather remains in the
follicle. For example, the concentric pinching of the
feather shaft, seen in psittacine circovirus infection, may
not be visualized until the feather is plucked from the fol-
licle. Look for stress bars in the wing and tail primaries
(Fig 26.1). Collect multiple blood feathers, both plucked
and in the follicle, along with any skin lesions (Fig 26.2)
and place them in formalin. Check for any signs of
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Table 26.3 | Necropsy Checklist

Owner’s Name Date of Necropsy
Animal’s Name Date of Death
Species Euthanasia Method
Age Sex Body Weight at Necropsy
Band/Microchip # Tattoo?

Organ(s) Normal Abnormal Description

Skin/Feathers
Beak/Oral Cavity/Tongue
Eyes/Ears/Conjunctivae
Sinuses/Choana/Nasal Cavity
Skeletal muscle/Bones/Joints
Liver/Gall Bladder, if present
Spleen
Thyroids/Parathyroids
Trachea/Lungs/Airsacs
Kidneys/Adrenals
Testes/Ovary/Oviduct
Crop/Esophagus
Proventriculus/Ventriculus
Duodenum/Pancreas
Jejunum/Ileum/Ceca, if present
Colon/Cloaca
Bursa/Thymus
Brain/Meninges
Spinal Cord/Vertebrae/Nerves
Bone/bone marrow
Middle & inner ear
Heart/Great Vessels
Gut contents wet mount results:
Gut contents dried smear results:
Organ impression smear results:
Tissues in formalin:
Tissues frozen:

trauma or bruising. In neonates, closely examine the
umbilicus for cleanliness and the adequacy of healing.
Examine the unfeathered portions of the legs and the
feet for poxvirus lesions, bumblefoot, herpesvirus podo-
dermatitis and self-mutilation (Figs 26.3, 26.4). Examine
the uropygial gland, found at the base of the tail in some
species, and collect it for histopathologic evaluation, as
this can be a site of chronic inflammation and neoplasia.
Evaluate the beak, both the external and the intraoral sur-
faces (Fig 26.5). Open the mouth. Look at and under the
tongue for abnormalities. Look in the choanal slit for
mucus and exudate and for blunting of the choanal papil-
lae (Fig 26.6). Salivary gland enlargement can occur at the
base of the tongue and can be due to hypovitaminosis A,
bacterial abscesses or, rarely, mycobacterial infections.
The nares and ear canals should be clear and free of
debris or exudate. Examine the conjunctivae and the
nictitating membranes. In Columbiformes, these tissues
can be collected for Chlamydophila diagnostics, as they
may contain elementary bodies.
The infraorbital sinuses should be opened as aseptically
as possible, and swabs or aspirates collected for cytology
and culture of bacteria, Mycoplasma and fungi (Figs
26.7, 26.8). Bacterial sinusitis is quite common in
psittacines, but also occurs in passerine species, and
caseous exudate is often seen (Fig 26.9).
In cockatiels (Nymphicus hollandicus) with “lockjaw,”
sinusitis and temporomandibulitis are common, as well
as myositis of the mandibular muscles. The mandible
and its attached muscles can be placed in formalin for
histopathology. In these cases, bacteria such as Borde-
tella avium, Enterococcus, Escherichia coli and Entero-
bacter may be isolated. It is important to indicate to the
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Fig 26.1 | Dystrophic feathers from a sulfur-crested cockatoo
(Cacatua galerita) infected with psittacine circovirus. Note the
concentric pinching of the feather shafts.
Fig 26.3 | Cutaneous pox lesions of the feet in a red-tailed
hawk (Buteo jamaicensis).
Fig 26.4 | Avascular necrosis of the distal toes of a black-
crowned night heron (Nycticorax nycticorax).
bacteriology laboratory that B. avium is suspected
because this organism is somewhat fastidious and
colonies may take longer to appear. Bordetella avium
also may cause tracheitis, bronchitis and pneumonia in
cockatiels and rarely in other psittacines.
Several Mycoplasma species have been implicated in
conjunctivitis and sinusitis in psittacines and passerines,
but these require special media for isolation and are
recovered uncommonly. In small passerines, cross-
sections of the nasal cavity and sinuses can be submitted
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665
Fig 26.2 | Dermal lymphosarcoma in a western screech owl
(Otus kennicottii).
Fig 26.5 | Cutaneous pox involving the commissures of the
mouth in a red-tailed hawk.
in formalin, decalcified if needed, and examined histo-
logically. Sometimes the nasal cavity epithelium may be
the only site of viral inclusions diagnostic for canarypox.
Cryptosporidial rhinitis and conjunctivitis also can be
diagnosed with this method.
COELOMIC CAVITY
The coelomic cavity examination is begun by placing the
body in dorsal recumbency, incising the skin of the
abdomen and peeling it back caudally over the abdomen
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Fig 26.6 | Normal choanal slit of a blue-
fronted Amazon parrot (Amazona aestiva).
Choanal slit (A), caudal palate containing
salivary glands (B), choanal papillae (P),
palatine beak (C).
Fig 26.7 | View of the avian skull with the
lateral wall of the infraorbital sinuses cut
away in a blue-fronted Amazon parrot.
Exudate may accumulate in the temporo-
mandibular joint (TMJ), especially in cock-
atiels. Ear (E).
Fig 26.8 | View of the infraorbital sinuses
in a blue-fronted Amazon parrot with the
lateral wall removed. Infraorbital sinuses
(IS), zygomatic or jugal arch (Z).

and cranially over the pectoral muscle mass. Assess the

condition of the pectoral muscle, as this is a good meas-
ure of weight loss (Fig 26.10). Notice whether subcuta-
neous fat is present or absent and whether there is any
bruising or edema. Grasp the sternum with thumb for-
ceps and slightly elevate, maintaining tension on the
abdominal wall. Make a transverse incision with the
scalpel blade just caudal to the edge of sternum, being
careful not to lacerate the liver. Remove the keel and
pectoral muscles in one piece by cutting through the
ribs and shoulder girdle with scissors or rongeurs.

Liver and Spleen

Assess the size, color and consistency of the liver. Hepatic
margins should be sharp and not extend beyond the
caudal edge of the keel in an adult bird (Figs 26.11-
26.13). Note whether a gall bladder is present or absent,
as not all species have one. Assess the condition of the
air sacs and coelomic surfaces that are visible. If
coelomic fluid is present, collect it with a sterile syringe
for analysis. Aseptically collect samples of air sacs next,
since they are delicate structures that readily disappear
with further manipulation of the organs. Aseptically col-
lect liver samples: one sample each for bacteriology,
virus isolation or DNA probe testing, Chlamydophila
testing and histopathology, and use any remaining tissue
for toxicology and/or impression smears.
Grasp the ventriculus, elevate and incise through the
attached membrane/air sac along the left margin and
rotate the ventriculus counterclockwise to find the spleen.
The spleen is nestled in the curve between the proven-
Fig 26.9 | Spirochetes in a wet mount of exudate from the
infraorbital sinuses of a cockatiel chick (Nymphicus hollandicus),
photographed under phase contrast.
triculus and the ventriculus (Fig 26.14). Evaluate the size
and shape of the spleen. Determining whether it is of
normal size for the bird being necropsied requires some
practice, so measuring the diameter can be helpful. The
spleen is round in some species, such as Psittaciformes
and Galliformes, and elongated or sock-like in Passeri-
formes and Columbiformes. Note the color and the pres-
ence of any pale foci in the spleen. Collect the entire
spleen, dividing it into three samples: one for virology,
one for Chlamydophila diagnostics and one for histo-
pathology. The spleen is the single most important sam-
ple for the histopathologic diagnosis of avian polyoma-
virus infection, since this is where viral inclusions are
most abundant.
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Fig 26.10 | View of the normal pectoral
muscles of a blue-fronted Amazon parrot
following removal of the skin. Crop (C).
Fig 26.11 | The liver extends beyond the
caudal edge of the keel, indicating hepato-
megaly in this mature red-tailed hawk with
multiple mycobacterial granulomas in the
liver.
Fig 26.12 | Marked hepatomegaly in a
Lady Gouldian finch (Chloebia gouldiae). In
this case, the hepatomegaly was due to lym-
phosarcoma, but a number of diseases can
cause marked hepatomegaly in passerines.

Fig 26.13 | Cholangiocarcinoma in an Amazon parrot Fig 26.14 | Normal spleen in a blue-fronted Amazon parrot.
(Amazona sp.). This tumor can have a very pleomorphic gross Spleen (S), liver (L), proventriculus (P), ventriculus (V).
appearance. Heart (H).

Genitourinary black) in some species, most notably cockatoos. Note

Reflect the ventriculus and the intestinal tract to the right
side of the bird to view the adrenals, gonads and kidneys,
leaving the unopened gastrointestinal tract for last to
avoid contamination of the other abdominal organs. The
adrenals are often obscured by active gonadal tissue, so it
is easier to collect the cranial division of the kidney with
the adrenal and gonad(s) attached for histopathology.
Adrenalitis is sometimes noted in unexplained death and
may be the only abnormality in some psittacines with
proventricular dilatation disease (PDD).
Sex the bird visually. In most species, only the left ovary
and oviduct develop in females, but both testes develop
in male birds. The gonads may be pigmented (brown or
the degree of development of the ovary and oviduct. Is
there follicular development? If so, record the general
size of the follicles. Is the oviduct hypertrophied?
Open the oviduct to look for exudate and tumors, and
collect samples for bacteriology and histopathology as
needed. Neoplasms of the oviduct may occur in the
mucosa or myometrium. Testicular tumors are common
in budgerigars (Melopsittacus undulatus) and ducks (Fig
26.15), but seasonal testicular enlargement also occurs.
In some species, such as in many passerines, this enlarge-
ment can be mistaken for neoplasia; histopathology can
usually distinguish between these two changes.
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Fig 26.15 | Seminoma of the testes in a mallard duck Fig 26.16 | Dissection displaying kidneys, reproductive
(Anas platyrhynchos). structures and lung from a normal blue-fronted Amazon
parrot. Lung (L), ovary, immature (O), cranial division (Cr),
middle division (M) caudal division (Ca) of the kidney,
salpinx or oviduct (S).

The kidneys are nestled in the renal fossae of the syn-

sacrum, with the lumbosacral plexus lying deep to the
caudal divisions of the kidney bilaterally (Fig 26.16). The
ureters run down the ventral surface of the kidneys. In
addition to the kidney/adrenal/gonad tissue collected as
described above for histopathology, aseptically collect
additional renal samples for virology, toxicology and bac-
teriology (if exudate is present).

In small birds (under 30 g), one can make an en bloc

excision of the kidneys still in situ within the synsacrum
and place this in formalin. After fixation, renal dissection
is easier and/or the synsacrum can be decalcified and
cross-sections of kidney together with bone can be cut.
After removal of the kidneys, evaluate the lumbosacral Fig 26.17 | View of the thoracic inlet, thorax and cranial
plexus, especially in cases of pelvic limb weakness or abdomen following removal of the keel and pectoral mus-
malfunction. Samples of these nerves can be collected in cles of a blue-fronted Amazon parrot. Thyroids (Th) are
located cranial to the heart on either side of the trachea
formalin for histopathologic evaluation.
(Tr), close to the carotid arteries. The parathyroids are
located at the caudal margin of the thyroids and are barely
visible in normal birds.
THORACIC INLET
Move to the thoracic inlet region. Identify the thyroids
and parathyroids, located cranial to the heart and adja-
ered. Parathyroid hypertrophy is usually the only gross
cent to the carotid arteries bilaterally, and collect them
pathologic lesion found in the hypocalcemia syndrome
for histopathology (Fig 26.17). Goitrous changes were
of African grey parrots (Psittacus erithacus).
once quite common in budgerigars, but are less so with
the advent of commercial diets. Hyperplastic goiter has
In young birds, multiple pale lobules of thymic tissue
been reported in juvenile macaws recently, but the cause
can be found along the cervical region, from the jaw to
is currently unknown. Lymphocytic thyroiditis also may
the thoracic inlet (Figs 26.18, 26.19). Thymic tissue can
be seen histologically, especially in Amazon parrots
be collected for virology and histopathology. Thymomas
(Amazona spp.). Thyroid tumors are uncommon, but
occasionally are seen as circumscribed, fairly encapsu-
seem to be observed most often in cockatiels, where
they tend to be very vascular. lated masses; they tend to be slow growing, undergoing
cystic degeneration with large areas of hemorrhage.
Normal parathyroids are barely visible. When they are Thymic tissue is usually quite difficult to detect in adult
prominent, metabolic bone disease should be consid- birds.
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Fig 26.18 | Left lateral view of

the neck of a normal young blue-
fronted Amazon parrot revealing
multiple lobules of thymic tissue
along its entire length from the jaw
to the thoracic inlet. Thymus (Th),
crop (Cr).
Fig 26.19 | Right lateral view of the neck
of a young blue-fronted Amazon parrot
revealing multiple lobules of thymic tissue
along its entire length from the jaw to the
thoracic inlet. Thymus (Th), trachea (Tr).
Fig 26.20 | Visceral gout in a red-tailed hawk
with severe deposition of urates in the pericar-
dial sac and in the perihepatic membranes. This
is a very important gross lesion for the practi-
tioner to recognize, because the urates may dis-
solve out of the tissue when placed in formalin.
A smear of the white material can be examined
under polarized light to reveal the typical urate
crystal structure. Trachea (T), liver (L), heart (H).

Cardiac Cardiomyopathy, usually the dilative form, can occur in

Examine the heart, pericardium and great vessels.
Visceral gout can cause the deposition of white, mucoid
urate material in the pericardial sac (Fig 26.20). View a
smear of this material under polarized light to confirm
the presence of uric acid crystals. It is very important to
recognize the gross appearance of visceral gout, because
formalin fixation may dissolve the uric acid crystals and
they may not be visible on histopathology (crystals do
not dissolve if fixed in ethanol, but this is usually not a
practical fixative for other reasons).
birds. The cause of the cardiomyopathy is often obscure
by the time it becomes a clinical problem, but myocar-
dial degeneration and fibrosis are often seen histologi-
cally. After removing the heart and great vessels, open
the heart in the direction of blood flow, using water to
rinse away blood and clots. (See Chapter 12, Evaluating
and Treating the Cardiovascular System for measure-
ments of the heart). Look for thrombi, valvular endo-
carditis lesions and pale areas in the myocardium.
Congenital cardiac anomalies are rarely diagnosed.

Suppurative pericarditis can be caused by a variety of
bacteria such as Pasteurella and Chlamydophila.
Cytologic examination of the pericardial exudate may
reveal the causative organisms. Portions of the pericar-
dial sac can be included in the tissues used for Chlamy-
dophila diagnostics. Hydropericardium is a common
finding in avian polyomavirus infection in juvenile
psittacines.
Open the great vessels to look for atherosclerosis, which
may involve the aorta, pulmonary artery or carotids.
Atherosclerosis is characterized grossly by yellowish,
raised, intimal plaques, but occasionally may be so
severe that the carotids are completely obstructed.
Mineralization of the great vessels also may occur in
association with atherosclerosis or may be related to
renal disease and hypervitaminosis D.

Prior to removing the heart from the thoracic cavity, heart
blood can be collected using a sterile syringe and needle
for bacteriology. Smears of heart blood can be stained
with Wright’s stain and examined for hemoprotozoa and
microfilaria, or Gram’s stained to look for bacteria.
Atherosclerosis is most commonly seen in African grey
parrots, where it can be mild to moderate, but also in
obese, older Amazon parrots, older macaw species (Ara
spp.) and captive raptors, where it can be so severe that
it results in acute death (Fig 26.21). Atherosclerotic
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Fig 26.21 | Atherosclerotic plaques on the luminal surfaces of
the great vessels of the heart in an Amazon parrot (Amazona
sp.). Note the smooth, glistening luminal surfaces of the normal
heart on the left.
lesions also may be found in the coronary arteries, but
usually these lesions are discovered upon histopathologic
examination. Atherosclerosis is a commonly missed diag-
nosis because the vessels are not opened and examined.
It is best to place most of the heart in formalin so that
multiple sections can be cut for histopathologic evalua-
tion. Pale foci or streaks can indicate degenerative
myopathy related to vitamin E/selenium deficiency, or
myocarditis associated with septicemia or viral diseases
such as West Nile virus or PDD. Petechial and ecchy-
motic epicardial hemorrhages are commonly seen in
cases of acute death from avian polyomavirus.
Lungs
Examine the lungs in situ prior to removing them. Avian
lungs are fixed in place within the avian thoracic cavity
and are not freely moveable. Removal requires gentle
teasing of the lung tissue away from the ribs. The avian
lung is one tissue in which gross lesions may appear
quite significant, but upon histopathologic evaluation
turn out to be just passive congestion. Conversely,
grossly normal lungs may contain significant histologic
lesions. So, it is wise to always include lung for histo-
pathology, even if it appears grossly normal. Collect a
portion of the lung for bacteriology and virology and
place the rest in formalin for histopathology. Because
lesions can be focal or multifocal, it is best to include a
large portion of at least one lung for histopathology.
In canaries and mynahs, a Wright’s-stained impression
smear of lung (along with impressions from liver and
spleen) is very important in detecting the monocytic
form of atoxoplasmosis, as Atoxoplasma organisms are
not usually visible on histopathology. Sarcocystis organ-
isms, bacteria and fungi also can be seen in impression
smears of the lung.
Page 670
ORAL CAVITY AND
GASTROINTESTINAL TRACT
Cutting through the mandible at one of the lateral com-
missures allows access to the caudal pharynx and visuali-
zation of the glottis (Figs 26.22, 26.23). The larynx and
trachea can then be opened down to the tracheal bifur-
cation, looking for hemorrhage, exudate, foreign bodies,
granulomas, and parasites such as respiratory mites or
Syngamus nematodes. Laryngeal papillomatosis can
occur in the pharynx and may occlude the glottis
(Fig 26.24).
Tissue samples should be collected for histopathology
and virus isolation, as well as bacteriology and fungal
culture if warranted. Fungal tracheitis, especially at the
syrinx, can be diagnosed by cytologic examination of
exudate or granulomatous material, fungal culture
and/or by histopathology. Rarely, mycobacterial organ-
isms can cause syringeal granulomas.
In canaries, Enterococcus faecalis can cause chronic
tracheobronchitis; culture of the tracheal lumen is nec-
essary for diagnosis. Viral tracheitis is rare in psittacine
birds, but a herpesviral tracheitis, bronchitis and airsac-
culitis have been reported in Neophema parrots. Canary-
pox can produce a severe tracheobronchitis with intracy-
toplasmic inclusions. A severe, chronic tracheobronchitis
can be seen with Bordetella avium in cockatiels; this
organism also is often associated with the “lockjaw” syn-
drome of sinusitis and temporomandibulitis.
Esophagus and Crop
Going back to the pharynx, the cut can extend down-
ward the length of the esophagus and into the crop,
looking for lacerations, punctures, peri-esophageal
abscesses and other abnormalities. In game birds, the
esophagus and crop may exhibit moderate to marked
thickening and mucus production due to capillariasis. A
wet mount scraping from the crop can reveal the typical
bipolar capillarid ova. In juvenile psittacines, thickening
and “Turkish towel” appearance of the crop mucosa is
often due to candidiasis, and either a wet mount smear,
cytology or Gram’s stain of a crop mucosal scraping can
be diagnostic (Fig 26.25).
Trichomonads can be found in wet mounts from the
oral cavity and/or crop of Columbiformes and raptors,
but also may occasionally be found in the crops of
budgerigars and passerines. The crop contents can be
collected in a plastic bag and frozen, if there is any sug-
gestion of toxin ingestion. A large section of crop, to
include a large vessel and adjacent nerve, should be col-
lected for histopathology, since PDD lesions are often
closely associated with the nerves.
26_Necropsy.qxd 8/23/2005 2:34 PM Page 671
Fig 26.22 | Cutting through the mandible allows opening of
the entire oral cavity and better visualization in a blue-fronted
Amazon parrot.
Fig 26.24 | The glottis of this Fig 26.25 | Severe crop mycosis
lilac-crowned Amazon parrot in a cockatiel chick.
(Amazona finschi) is occluded by
laryngeal papillomatosis.
At this point, the esophagus distal to the crop can be
transected. Caudal traction of the distal esophagus and
sharp dissection of the mesenteric attachments can be
utilized to remove the entire gastrointestinal tract.
Bursa of Fabricius
Continue the dissection to make a circular incision
around the vent, leaving a margin of intact vent skin and
the bursa of Fabricius attached to the tract. The bursa is
present in young birds usually less than 6 to 12 months
of age and is located dorsal to the cloaca. The bursa
should always be collected when it is present and
divided in half. Submit one half in formalin for histo-
pathology and save the other half for virology and/or
DNA probe testing.
Chapter 26 | D I A G N O S T I C V A L U E O F N E C R O P S Y
671
Fig 26.23 | Normal glottis from a blue-fronted Amazon parrot.
Tongue (T), glottis (G).
The bursa is important in diagnosing psittacine cir-
covirus, especially in young African grey parrots that
die acutely without feather lesions, since the bursa may
be the only site where viral inclusions are found.
Circoviral inclusions also can be found in the bursa of
young pigeon squabs dying from a variety of secondary
infections. Lesions in the bursa are often non-specific
as to etiology, but can indicate the acuteness or
chronicity of stress.
Proventriculus and Ventriculus
Open the distal esophagus with scissors, continuing
on into the proventriculus and ventriculus. Evaluate
the stomach contents for amount and any foreign
material. Washing the contents into a bowl or strainer
can allow the food material to be rinsed away, leaving
metallic and other foreign bodies behind. Collect and
freeze the contents for possible toxicologic analysis.
Rinse the mucosa with water and make wet mount
and dried smears of mucus and/or mucosal scrapings.
Do not separate the proventriculus and ventriculus.
The isthmus (the junction between the proventriculus
and ventriculus) is a common site for avian gastric
yeast (formerly known as megabacteria); its suggested
new name is Macrorhabdus ornithogaster (see
Chapter 30, Implications of Macrorhabdus in Clinical
Disorders) (Fig 26.26). Grey-cheeked parakeets (Broto-
geris pyrrhopterus) seem to be prone to the develop-
ment of gastric carcinoma at the isthmus and the gross
lesions are often unexciting. Collect a large specimen
of proventriculus, isthmus and ventriculus (all in one
piece if possible), containing at least one large serosal
nerve and blood vessel, for histopathology.
In small birds, the entire proventriculus and ventricu-
lus can be placed in formalin. A large specimen
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672 C l i n i c a l Av i a n M e d i c i n e - Vo l u m e I I
Fig 26.26 | Large gram-positive organisms consistent with the
yeast Macrorhabdus ornithogaster (formerly known as mega-
bacteria) are seen in a Gram’s-stained smear of intestinal con-
tents from a cockatiel. Note the size difference between these
fungal organisms and the smaller gram-positive bacilli.
allows multiple sections to be examined by the veteri-
nary pathologist in the search for nerves and plexi.
Dilatation of the proventriculus and/or ventriculus is a
hallmark gross lesion of PDD, but in juvenile psittacines
being hand-fed, these organs also may be dilated as a
normal finding. Histopathology is required to differenti-
ate between PDD and normal juvenile underdevelop-
ment of the proventriculus and ventriculus (Fig 26.27).
Foreign body penetration of the ventricular wall can
occur in any species, but is most common in waterfowl
and ratites. Nutritional muscular dystrophy (degenera-
tive myopathy) can be seen in some species as white
streaks in the ventricular muscle as a manifestation of
vitamin E/selenium deficiency. Endoventricular mycosis
(fungal invasion of the koilin lining of the ventriculus)
can be seen histologically and is a common finding in
debilitated passerines, despite the usually unremarkable
gross appearance.
Duodenum and Pancreas
Open the outflow tract from the ventriculus and pro-
ceed into the duodenal loop. The largest limb of the
pancreas lies in the duodenal loop mesentery while the
small splenic lobe of the pancreas is located adjacent to
the spleen. Pancreatic lesions are fairly common histo-
logically, but gross lesions may not be very striking. The
pancreas also is one of the first organs to undergo post-
mortem autolysis.
Quaker parrots (Myiopsitta monachus) are prone to the
development of acute pancreatic necrosis of unknown
etiology. Fat necrosis and serositis may accompany pan-
creatitis and pancreatic necrosis. Quaker parrots that sur-
Page 672
Fig 26.27 | Marked dilatation of the proventriculus (P) in a
macaw (Ara ararauna) with proventricular dilatation disease
(PDD). H= heart, V= ventriculus.
vive the initial insult may develop severe pancreatic atro-
phy and fibrosis. Inclusion body pancreatitis can be seen
with herpesvirus and adenovirus infections. Lympho-
plasmacytic pancreatitis in Neophema parrots is associ-
ated with paramyxovirus infection. Pancreatic necrosis
also is a common lesion in West Nile virus infection.
Vacuolar changes and necrosis of acinar cells may be
seen in zinc toxicosis, but these lesions can be readily
obscured by even mild postmortem autolysis. The pan-
creas concentrates zinc in the acinar cells and should be
collected for toxicologic analysis, along with liver and
kidney, to diagnose zinc toxicity. Collect a sample of pan-
creas for virology. Also submit in formalin a transverse
section through the duodenal loop with pancreas
attached, as this helps to identify the duodenum.
Yolk Sac
In neonate, the yolk sac and stalk should be evaluated
for the degree of absorption. In psittacine and passerine
chicks, the yolk sac is usually quite tiny by 3 days after
hatching. Collect a sterile sample of the yolk material for
culture and place the rest of the yolk sac (wall and con-
tents) into formalin. Yolk sacculitis and yolk sac reten-
tion are common problems in neonatal ratites.
Intestines
Continue opening the intestine through the jejunum
and ileum to the ceca (if present in the species) and
colon. Collect sections of intestine for histopathology.
Opened intestinal sections are usually best, as this gives
the mucosa a chance to fix rapidly (Fig 26.28). Do not
disturb the mucosa by scraping or handling, as artifacts
26_Necropsy.qxd 8/23/2005 2:35 PM Page 673
Fig 26.28 | Intestinal volvulus in an ostrich (Struthio camelus).
can confuse or obliterate the histologic diagnosis.
Wet mounts of intestinal contents (usually two different
sites) are helpful in diagnosing parasitic and bacterial
problems. Wet mounts should be examined for parasite
ova and oocysts, as well as flagellates, yeast and motile
bacteria. Sections of bowel can be tied off with string or
suture and submitted for culture. In some cases, both aer-
obic and anaerobic culture may be warranted (Fig 26.29).
If necrotic lesions are encountered in the intestinal
mucosa, clostridial disease should be considered. Quail
disease caused by Clostridium colinum is a common
problem in quail, and typical “button ulcers” can be
seen in the intestines as well as “crateriform” necrotic
lesions in the liver. Clostridial enteritis, usually caused
by Clostridium perfringens, is becoming more com-
monly recognized in psittacines, especially nectar eaters
such as lories and lorikeets. Clostridial organisms in
large numbers can cause acute necrohemorrhagic enteri-
tis. Finding large gram-positive bacilli, with or without
spore formation, as the primary organism on a Gram’s-
stained smear of intestinal contents gives a presumptive
diagnosis that should be followed by anaerobic culture
(Fig 26.30). Because exposure to oxygen in the air can
inhibit clostridia, it is wise to tie off a loop of unopened,
affected bowel with string or suture and place it in a
sealable bag with the air evacuated prior to sending it
for anaerobic culture.
A wide variety of other bacteria can cause enteritis and
septicemia. Gram-negative organisms, especially Entero-
Chapter 26 | D I A G N O S T I C V A L U E O F N E C R O P S Y
673
Fig 26.29 | Fibrino-suppurative enteritis in a tufted puffin
(Fratercula cirrhata).
bacteriaceae, are common infectious agents in psittacines
and passerines. In addition, Campylobacter spp. and
Yersinia pseudotuberculosis are more common in
canaries and exotic finches. Campylobacter organisms
usually require special media and microaerophilic incu-
bation conditions, so it is wise to alert the bacteriology
laboratory when this organism is suspected.
Multifocal granulomas or thickened areas of bowel can
be indicative of mycobacteriosis. These sites should be
collected for histopathology, and special acid-fast tissue
stains can be applied to paraffin sections to demonstrate
the organisms. Alternatively, impressions or scrapings
from these sites can be stained with a rapid acid-fast stain
for a quick, presumptive diagnosis. Sections of affected
bowel can be collected for mycobacterial culture.
Intestinal neoplasia is fortunately uncommon in birds,
but needs to be included in the differential diagnosis of
thickened or proliferative bowel lesions.
Flagellate protozoa and coccidial organisms also may
produce enteritis. Flagellates (including Giardia spp.
and Cochlosoma spp.) are diagnosed by fresh wet
mount smears of intestinal contents, but they are nearly
impossible to diagnose on histopathology. Coccidiosis
can be diagnosed by wet mount smears of intestinal
scrapings and by histopathology. Nematode and cestode
parasites are uncommon in domestically raised
psittacines and passerines, but geographic pockets of
these parasites may exist and should always be consid-
ered (Fig 26.31). These parasites are still common in
ground-feeding and feral or wild birds.
Ceca
Many species of birds do not possess ceca. Psittacines do
not. Passerines and Columbiformes have tiny vestigial
ceca composed of lymphoid tissue, while Galliformes,
Anseriformes and ratites possess large bilateral ceca.
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674 C l i n i c a l Av i a n M e d i c i n e - Vo l u m e I I
Fig 26.30 | A Gram’s-stained smear of intestinal contents from
a lory (Trichoglossus haematodus) that died of acute necrohem-
orrhagic enteritis. Clostridium perfringens was isolated from the
intestinal tract. Note the large gram-positive rods with sub-
terminal spores.
These should be opened to look for cecal worms and
their contents should be included for culture. Cecal con-
tents should be included in culture for Salmonella,
especially in Galliformes.
Colon
Colon contents should start to look like fecal material as
one moves toward the cloaca. Open the cloaca to look
for papillomatous lesions, cloacoliths, trauma, inflamma-
tory lesions and neoplasia (Fig 26.32).
In summary, intestinal samples should include the fol-
lowing: wet mounts from at least two different sites,
smears for Gram’s stain and possibly acid-fast stain, con-
tents for aerobic and possibly anaerobic bacteria or
Campylobacter culture, tissue for histopathology and
ingesta for virology (direct electromicroscopy, virus isola-
tion and/or DNA probes), and toxicology.
NEUROLOGIC
The brain and spinal cord can be very important in the
diagnosis of some diseases, especially PDD. The dorsal
calvarium should be carefully removed with rongeurs
(Fig 26.33). Visualize the brain in situ for any obvious
abnormalities such as abscesses, which should be cul-
tured. Remove the brain by inverting the skull and tran-
secting the ventral and cranial attachments (Fig 26.34).
Collect a portion of the forebrain for virology and toxi-
cology, and fix the rest of the brain in formalin. In
neonates, the brain is so soft that making a cut through
the dorsal skull and placing the entire calvarium con-
taining the brain in situ into formalin is recommended.
After fixation, the brain will harden somewhat and it can
be removed more easily without damaging it.
A similar procedure can be followed for the cervical
Page 674
Fig 26.31 | Tapeworms attached to the intestinal mucosa of a
great horned owl (Bubo virginianus).
Fig 26.32 | Cloacal papillomatosis is seen in a lilac-crowned
Amazon parrot. The cloaca has been opened caudally to cra-
nially.
spinal cord. Cut the vertebral column with cord in situ
into 2- to 3-cm pieces and fix in formalin overnight. This
process will allow easier removal using rongeurs, with
minimal damage to the less fragile, fixed spinal cord. In
very small birds, cross-sections of the cervical vertebral
column with the spinal cord in situ can be decalcified
and examined histologically.
In birds with head tilt or neurologic disease, especially
Neophema parrots and exotic finches, fix a large portion
of the petrous temporal bone containing the middle ear.
This bone can later be decalcified by the veterinary
pathologist and sectioned to examine the middle ear for
inflammation and viral inclusions associated with
paramyxovirus infections. Congestion of the vascular
sinuses in the bones of the skull is a common finding,
but it is significant only if there also is corresponding
subdural hemorrhage or bleeding of brain parenchyma.
The eyes can be removed and fixed in formalin if there
is any suspicion of blindness, ocular or neurologic dis-
ease. Dissection through some orbital bone may be
required for removal. Remember that the avian eye has
bony scleral ossicles, which may make their sectioning
somewhat more difficult. The globes can be transected
at the optic nerves or dissection can be carried out
26_Necropsy.qxd 8/23/2005 2:35 PM Page 675
Fig 26.33 | View of the dorsal surface of calvarium of a
psittacine bird with congested vascular sinuses (S).
Fig 26.35 | View of the ventral surface of the brain from a
blue-fronted Amazon parrot. Pituitary (P), optic lobes (O), brain-
stem (BS), optic chiasm (C), optic nerves (N).
through the ventral calvarium to keep the optic nerves
and chiasm intact and attached to the brain. On the ven-
tral surface of the brain near the optic chiasm is the
pituitary (Fig 26.35). Tumors of the pituitary have been
reported in budgerigars and cockatiels (Fig 26.36).
MUSCULOSKELETAL
Bone marrow can be collected by aspiration of the
femur and smears made and stained for cytologic evalua-
tion. Collect a segment of femur using rongeurs and
place it in formalin. Once fixed, the previously fragile
bone marrow can be dissected out and examined histo-
logically. Leukemic or aplastic processes can be diag-
nosed from bone marrow samples, and circovirus inclu-
sions also may occasionally be seen histologically.
Samples of skeletal muscle should be collected for
histopathology. Muscular lesions may include trauma,
hemorrhage, degeneration, mineralization, and injection
or vaccine site reactions. Myositis, degenerative myopa-
Chapter 26 | D I A G N O S T I C V A L U E O F N E C R O P S Y
675
Fig 26.34 | Dorsal surface of a normal brain from a blue-
fronted Amazon parrot.
Fig 26.36 | A pituitary tumor in a budgerigar (Melopsittacus
undulatus).
thy and Sarcocystis infection can be diagnosed histologi-
cally (Fig 26.37). Open the joints of the pelvic and tho-
racic limbs and look for exudate; collect synovial fluid
with a sterile syringe for bacterial and mycoplasmal cul-
ture, although exudate also can be caseous.
Articular gout can be diagnosed by examining the exu-
date on cytology or by histopathology. The lesions of
degenerative joint disease, periarticular proliferation and
proliferative synovitis are fairly common in the joints of
the feet, but also may occur in the shoulder, stifle and
hock. Any bone or joint lesions demonstrated radi-
ographically should be opened and sampled for culture
and histopathology (Fig 26.38).
The flexibility of bones (eg, tibiotarsus, ribs) can be used
in the assessment of the adequacy of mineralization. The
bones should break with an audible snap if mineraliza-
tion is normal. The rachitic “rosary” at the costochondral
or costovertebral junctions and deformation of the keel
or other long bones are obvious lesions of metabolic
26_Necropsy.qxd 8/23/2005 2:35 PM
676 C l i n i c a l Av i a n M e d i c i n e - Vo l u m e I I
Fig 26.37 | Pale areas in skeletal muscle from an emu
(Dromiceius novaehollandiae) with degenerative myopathy. The
heart muscle appears grossly normal.
bone disease. Sections of bone, especially areas of the
metaphyses and epiphyses, can be examined histologi-
cally for metabolic bone disease.
Ratites are prone to developing angular limb deformities.
The “rubber rhea” syndrome is often due to hypophos-
phatemic rickets. Other ratite limb deformities can be
multifactorial, but nutritional imbalances in calcium,
phosphorus and vitamin D3, growth rates and problems
with substrates are often implicated. Flock problems with
limb deformities in ratites can be investigated through
feed/forage analysis and bone ash analysis.
SMALL BIRDS AND DEAD-IN-SHELL
Necropsy of very small birds and neonates (under 15 g)
is challenging. One can open the coelomic cavity and
thorax and fix the entire body in formalin; opening the
ventriculus as well is recommended for best fixation.
The veterinary pathologist can then carry out dissection.
It is very difficult to get good fixation of tissues in birds
weighing more than 20 g, so this technique should not
be used for them.
Dead-in-shell and egg necropsies can be performed, but
there are limited lesions and testing available. Open the
egg as aseptically as possible at the air cell end. Collect
samples aseptically for bacterial culture, virology and
DNA probes. Assess gestational age and positioning of
the embryo or chick. Then place the embryos and mem-
branes in formalin. Histopathologic diagnosis is often
limited by autolysis, since the eggs often remain in the
incubator for a period of time after embryonic death.
Histologic examination of the blastodisk can help deter-
mine if the eggs are truly infertile or whether early
embryonic death occurred post-fertilization.
Page 676
Fig 26.38 | A proliferative osteosarcoma involving the right leg
and pelvis of a peach-faced lovebird (Agapornis roseicollis).
Ancillary Testing of
Samples Collected at
Necropsy
After the necropsy has been concluded, the remaining
parts of the carcass can be placed in a sealable plastic
bag and frozen until diagnostic testing has been com-
pleted. Examine wet mounts of intestinal contents and
crop or oral cavity scrapings as quickly as possible in-
house for parasite ova, oocysts, motile flagellates, yeast
and motile bacteria.
MICROBIOLOGY
Stain impression smears from organs and bone marrow,
smears of exudate, or cells from fluid analysis with a
Wright’s stain and examine for cell types and microor-
ganisms, including bacteria and fungi. If bacteria are
seen on the cytologic preparations, a stained smear can
be destained and restained with Gram’s stain, or another
smear can be stained. If macrophages with “ghost
bacilli” are seen on the cytologic preparation, an acid-
fast stain is in order to attempt to demonstrate myco-
bacterial organisms.
A Gram’s stain of the colonic contents can be performed
in-house to provide a quick evaluation of the presence
of abnormal bacterial populations, and then followed up
with culture. This type of Gram’s stain is especially
important in detecting possible clostridial organisms,
which would then prompt an anaerobic culture. Avian
gastric yeast also can be detected in fecal smears or from
scrapings of the isthmus stained with cytologic stains.
Composite Samples
A collection of liver, spleen and air sac can be submitted
26_Necropsy.qxd 8/23/2005 2:35 PM Page 677
for Chlamydophila diagnostics. A Gimenez or Macchia-
vello stain can be performed on impression smears of air
sac, liver and spleen for the demonstration of elemen-
tary bodies. In Columbiformes, conjunctiva and nictitat-
ing membrane should be included, as elementary bodies
may be confined to this location in these species. Fluore-
scent antibody and chlamydial culture may be available
at certain laboratories, and some also can perform a
DNA probe for Chlamydophila on a swab from the com-
bined surfaces of liver, air sac and spleen.
Tissues, exudates or swabs can be submitted to diagnos-
tic laboratories for bacterial, mycoplasmal or fungal cul-
ture as indicated. With the exception of samples for
Campylobacter, which does not survive freezing well,
these samples can often be frozen if not sent for culture
immediately.
Special media is required for the culture of Mycoplasma
spp. Alert the bacteriology laboratory if fastidious organ-
isms such as Campylobacter spp. and Bordetella avium
are of interest in the particular species or individual bird,
as these organisms often require special media and incu-
bation parameters. DNA probe testing for Salmonella
spp. is available at some laboratories. An antibiotic sensi-
tivity tailored to drugs used in pet avian species also can
be requested if other birds on the premises are at risk.
Mycobacterial culture is required for accurate speciation
of acid-fast organisms, and special media and handling
are necessary. Once Mycobacterium isolates are grown
on solid media, some laboratories are capable of speciat-
ing the organisms by the use of DNA probes and may
offer antibiotic sensitivity testing for mycobacterial
isolates.
Fungal culture may be requested in cases of suspected
mycoses and is often required for accurate identification
of the species involved. Antifungal sensitivity testing is
available at specialized mycology laboratories.
A pool of parenchymal tissues (liver, spleen, air sac,
lung, kidney, brain and bursa if present) and a separate
pool of intestinal contents should be refrigerated or
frozen for possible virus isolation or DNA probe testing.
A combination swab from heart blood and the cut sur-
faces of liver, spleen, lung, kidney and bursa can be
submitted for DNA probe testing for viruses such as
psittacine circovirus and avian polyomavirus. Fluorescent
antibody techniques on frozen sections of tissue may be
available for certain viruses.
Polymerase chain reaction (PCR) tests are available for
the detection of certain viruses, such as West Nile virus,
on fresh or frozen tissues. It is important to contact the
individual laboratory so the most appropriate tissues are
Chapter 26 | D I A G N O S T I C V A L U E O F N E C R O P S Y
677
submitted, as this can depend on the particular virus or
antigen and upon the particular laboratory’s technique.
PARASITOLOGY
Direct wet mounts of intestinal contents and crop or
oral cavity scrapings prepared and examined at the time
of necropsy are invaluable in the diagnosis of trichomo-
niasis in pigeons, raptors and budgerigars; cochlosomia-
sis in finches and canaries; and giardiasis in cockatiels
and other psittacine and passerine species. Many of
these organisms dry up easily, so examination should be
performed promptly. Examination of these wet mounts
under dark field or phase contrast, if available, may
make the detection of flagellates and motile bacteria eas-
ier. Inoculating Diamond’s media and submitting the
media for incubation can attempt culture of some tri-
chomonad parasites.
Microscopy of whole parasites such as nematodes, ces-
todes, flukes and acanthocephalans may provide mor-
phology that can point to the classification of the para-
sites, plus characteristic ova may be visible within the
helminths. An acid-fast or auramine stain can be per-
formed on smears for the detection of Cryptosporidium
oocysts, which can be found in the intestine, conjunc-
tiva, nasal cavity or bursa.
A stained smear of the heart blood or lung impression
can be examined for microfilaria and hematozoa such
as Plasmodium, Hemoproteus and Leucocytozoon.
Wright’s-stained impression smears of lung, spleen and
liver are especially important in canaries and finches for
the diagnosis of the monocytic form of atoxoplasmosis,
as these organisms may not be visible histologically.
Rarely, flagellates can be demonstrated in impression
smears from the lung, trachea, sinus and conjunctiva,
which are not readily visible histologically. Other proto-
zoal parasites such as Sarcocystis, Toxoplasma and Leuco-
cytozoon can be found in impression smears of organs.
HISTOPATHOLOGY
Select a group of formalin-fixed tissues with lesions or
a group of tissues that commonly contain histologic
lesions that could lead to diagnosis and submit them for
histopathology. This commonly includes tissues such as
liver, spleen, air sac, kidney, lung, trachea, heart, bursa,
brain, duodenum/pancreas and proventriculus/ventricu-
lus. Save the remaining formalin-fixed tissues in case the
diagnosis is not made with the first set of tissues.
This second set of tissues may include spinal cord, bone
marrow, nasal cavity, skin and feathers, bone and joint,
middle and inner ear, eyes, tongue, skeletal muscle, thy-
roid, parathyroid, adrenal, esophagus, crop, jejunum,
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678 C l i n i c a l Av i a n M e d i c i n e - Vo l u m e I I
ileum, colon, ceca, gall bladder, ovary, oviduct, testes,
thymus, nerve (ischiatic, brachial plexus) and beak.
The veterinary pathologist may recommend special diag-
nostics such as stains for acid-fast organisms, fungi, bac-
teria, iron or copper, depending on what is seen on the
routine hematoxylin- and eosin-stained sections. In spe-
cial situations, tissues may be embedded in plastic so
that electron microscopy can be performed. Direct elec-
tromicroscopy also can be performed on intestinal con-
tents or tissue homogenates. In situ DNA hybridization
techniques on paraffin-embedded tissues are available
for certain viruses such as Pacheco’s herpesvirus, aden-
ovirus, avian polyomavirus, psittacine circovirus and
paramyxovirus.
Immunohistochemical stains can detect certain antigens
from bacteria, fungi, viruses and parasites in paraffin-
embedded tissues, and these techniques also can be uti-
lized to detect some cell markers in the diagnosis of
tumors. Gene sequencing of certain microorganisms
(Clostridium perfringens, for example) in formalin-fixed,
paraffin-embedded tissues is available at some diagnostic
laboratories.
TOXICOLOGY
Toxicologic testing requires some idea of what toxin is
being considered. This information often comes from
the history and histopathologic findings. Contacting the
toxicology laboratory is essential for submission of the
most appropriate tissues and amounts.
The most common toxins tested for are heavy metals
such as lead and zinc. Usually liver and kidney are
required for this analysis, although zinc also accumulates
in the pancreas preferentially. Heavy metals also can be
detected in foreign bodies, water and feed. Copper accu-
mulation in the livers of swans can be demonstrated
qualitatively with special histologic stains for copper, but
quantitative levels require toxicologic analysis.
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Iron storage disease is most commonly seen in toucans,
toucanettes, mynahs and birds of paradise; the condition
is rare in psittacines, although there is emerging evidence
that lories and lorikeets may be prone to iron accumula-
tion. The special histologic stain, Perl’s Prussian blue,
can provide qualitative information about the amount of
iron in the liver, but, again, quantitative levels are
detected by toxicologic analysis.
Poisonous plants can be found in the digestive tract and
submitted to a botanist or university botany department
for identification. The plants or wood can be frozen
until submission to prevent the breakdown of toxic prin-
ciples. Ingestion of fertilized plants can result in nitrate
toxicity, and samples of these plants can be analyzed for
the amount of nitrates present.
Polytetrafluoroethylene (PTFE or non-stick coatings) and
other toxic inhalation products are rarely detectable in
tissues, and the diagnosis is usually made by a history of
exposure, the presence of pulmonary edema and hemor-
rhage, and the exclusion of other causes of death. There
is a wide variety of items commonly found in the house-
hold that can give off PTFE fumes, including non-stick
cookware and appliances such as self-cleaning ovens and
electric grills.
Birds also can be sensitive to other inhalants such as car-
bon dioxide, carbon monoxide and fumes from glues,
resins, plastics and paints. Mycotoxins may be implicated
in the case of multiple birds suffering liver damage.
Aflatoxins can be detected in foodstuffs, but usually by
the time chronic liver damage is evident, the offending
foodstuff is often no longer available. In the case of
acute toxicosis, samples of the feed should be frozen
along with liver and kidney, pending further investiga-
tion. Contact the toxicology laboratory for specimen
requirements and costs.